Method of producing antitumor effect using quinazolin-4-one derivatives

ABSTRACT

A quinazoline of the formula: ##STR1## wherein R 1  is alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, alkylthio, aryl, aryloxy, arylalkyl, halogeno, hydroxy, mercapto, pyridylthio, pyrimidinylthio, or substituted alkyl or alkoxy; 
     wherein R 2  is hydrogen, alkyl, alkenyl, alkynyl, substituted alkyl or alkanoyl; 
     wherein Ar is phenylene, naphthylene or heterocyclene which is unsubstituted or bears one or more substituents and wherein R 3  is such that R 3  --NH 2  is an amino acid; 
     or a pharmaceutically-acceptable salt or ester thereof. The compounds possess anti-tumor activity.

This is a division of application Ser. No. 07/334,748, filed Apr. 6,1989, now U.S. Pat. No. 4,992,550, which is a continuation of Ser. No.07/030,424, filed Mar. 26, 1987, now abandoned.

This invention relates to novel anti-tumour agents and more particularlyit relates to quinazoline derivatives which possess anti-tumouractivity.

One group of anti-tumour agents comprises the antimetabolites which areantagonists of folic acid, such as aminopterin and methotrexate. A newercompound of this type which showed considerable promise in clinicaltrials is known as CB3717 and is described and claimed in United KingdomPatent Specification No. 2065653B. Despite its promise, however, CB3717shows symptoms of toxicity in humans, particularly in relation to theliver and kidney.

Compounds of this type are believed to act as anti-tumour agents byinhibiting the enzyme thymidylate synthetase. Their anti-tumour activitymay be assessed in vitro by determining their inhibitory effect on thatenzyme, and in cell cultures by their inhibitory effect on the cell lineL1210.

We have now found that certain quinazoline derivatives are considerablymore active than CB3717, and furthermore are more water-soluble thanthat compound, which may be clinically important by increasing the easeof clearance through the kidney thereby decreasing any symptoms oftoxicity.

According to the invention there is provided a quinazoline of theformula: ##STR2## wherein R¹ is alkyl, cycloalkyl, alkenyl, alkynyl,alkoxy or alkylthio each of up to 6 carbon atoms;

aryl, aryloxy or arylalkyl each of up to 10 carbon atoms; halogeno,hydroxy, mercapto, pyridylthio or pyrimidinylthio; alkyl of up to 3carbon atoms which bears one or more substituents selected fromhalogeno, hydroxy, amino, pyridylthio, pyrimidinylthio, alkoxy,alkanoyloxy, alkylthio, alkylamino, dialkylamino and alkanoylamino eachof up to 6 carbon atoms and aroyloxy and aroylamino each of up to 10carbon atoms;

or alkoxy of up to 3 carbon atoms which bears one or more substituentsselected from hydroxy and alkoxy of up to 6 carbon atoms;

wherein R² is hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkyl,alkoxyalkyl, mercaptoalkyl, alkylthioalkyl, halogenoalkyl, cyanoalkyl,aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkanoylalkyl,carboxyalkyl, carbamoylalkyl or alkanoyl each of up to 6 carbon atoms oraroylalkyl of up to 10 carbon atoms;

wherein Ar is phenylene, naphthylene or heterocyclene which isunsubstituted or which bears one or more substituents selected fromhalogeno, phenyl, cyano, nitro, hydroxy, amino and carbamoyl and alkyl,alkoxy, halogenoalkyl, alkanoylamino, alkylthio and alkoxycarbonyl eachof up to 6 carbon atoms; and

wherein R³ is such that R³ --NH₂ is an amino acid; or apharmaceutically-acceptable salt or ester thereof.

A suitable value for R¹ or R² when it is alkyl or for an alkylsubstituent in Ar is, for example, methyl, ethyl, propyl, isopropyl,isobutyl, tert-butyl, pentyl or hexyl.

A suitable value for R¹ when it is cycloalkyl is, for example,cyclopropyl, cyclopentyl or cyclohexyl.

A suitable value for R¹ or R² when it is alkenyl is, for example,prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl,hex-5-enyl or 2,3-dimethylbut-2-enyl.

A suitable value for R¹ or R² when it is alkynyl is, for example,prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl,hex-2-ynyl or hex-5-ynyl.

A suitable value for R¹ when it is alkoxy, alkylthio or for an alkoxy oralkylthio substituent in Ar is, for example, methoxy, ethoxy,isopropoxy, hexyloxy, methylthio, isopropylthio or hexylthio.

A suitable value for R¹ when it is aryl or arylalkyl is, for example,phenyl, tolyl, benzyl, α-methylbenzyl or phenethyl.

A suitable value for R¹ when it is aryloxy is, for example, phenoxy ortolyloxy.

A suitable value for R¹ when it is halogeno is, for example, fluoro,chloro, bromo or iodo.

A suitable value for R¹ when it is substituted alkyl is, for example,fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl,3-fluoropropyl, chloromethyl, dichloromethyl, hydroxymethyl,2-hydroxyethyl, 3-hydroxypropyl, aminomethyl, 3-aminopropyl,pyrid-2-ylthiomethyl, pyrimidin-2-ylthiomethyl, methoxymethyl,isopropoxymethyl, 3-methoxypropyl, acetoxymethyl, propionyloxymethyl,methylthiomethyl, 3-methylthiopropyl, propylthiomethyl,methylaminomethyl, propylaminomethyl, methylaminopropyl,dimethylaminomethyl, diethylaminomethyl, ethylmethylaminomethyl,3-dimethylaminopropyl, acetamidomethyl, 3-acetamidopropyl,propionamidomethyl, benzoyloxymethyl or benzamidomethyl.

A suitable value for R¹ when it is substituted alkoxy is, for example,2-hydroxyethoxy, 4-hydroxybutoxy, 3-hydroxy-2-methylpropoxy,2-methoxyethoxy, 3-methoxypropoxy or 2-ethoxyethoxy.

A suitable value for R² when it is hydroxyalkyl, alkoxyalkyl,mercaptoalkyl or alkylthioalkyl is, for example, 2-hydroxyethyl,3-hydroxypropyl, 2-methoxyethyl, 2-ethoxyethyl, 3-methoxypropyl,2-methoxypropyl, 2-mercaptoethyl, 3-mercaptopropyl, 2-methylthioethyl,3-methylthiopropyl or 2-ethylthioethyl.

A suitable value for R² when it is halogenoalkyl, cyanoalkyl,aminoalkyl, alkylaminoalkyl or dialkylaminoalkyl is, for example,2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 3-fluoropropyl,3-chloropropyl, cyanomethyl, 2-cyanoethyl, 3-cyanopropyl, 2-aminoethyl,3-aminopropyl, 3-amino-2-methylpropyl, 2-methylaminoethyl,2-dimethylaminoethyl, 2-ethylaminoethyl, 2-diethylaminoethyl,3-methylaminopropyl or 3-dimethylaminopropyl.

A suitable value for R² when it is alkanoylalkyl, carboxyalkyl,carbamoylalkyl or alkanoyl is, for example, acetonyl, 2-acetylethyl,propionylmethyl, 2-propionylethyl, 3-acetylpropyl, 4-acetylbutyl,carboxymethyl, 2-carboxyethyl, carbamoylmethyl, acetyl, propionyl orbutyryl.

A suitable value for R² when it is aroylalkyl is, for example, phenacylor 2-benzoylethyl.

A suitable value for Ar when it is heterocyclene is, for example, a5-membered or 6-membered aromatic (that is, fully unsaturated)heterocyclene diradical which contains up to 2 heteroatoms selected fromthe group consisting of oxygen, nitrogen and sulphur, for example,thienylene, pyridylene, pyrimidinylene, thiazolylene or oxazolylene.

A suitable halogeno, halogenoalkyl, alkanoylamino or alkoxycarbonylsubstituent in Ar is, for example, fluoro, chloro, bromo, iodo,fluoromethyl, difluoromethyl, trifluoromethyl, acetamido, propionamido,isopropionamido, methoxycarbonyl, ethoxycarbonyl or isobutoxycarbonyl.

A suitable value for R³ is such that R³ --NH₂ is a naturally-occurringamino-acid such as L-aspartic acid, L-glutamic acid, L-alanine,L-phenylalanine, L-serine, glycine or L-ornithine. Alternatively R³ maybe such that R³ --NH₂ is L-2-aminobutyric acid or a poly-L-glutamicacid. R³ may therefore have, for example, the formula: ##STR3## whereinm is an integer from 1 to 10, or the formula: ##STR4##

A suitable pharmaceutically-acceptable salt of a quinazoline of theinvention is, for example, an acid addition salt with, for example,inorganic or organic acids, for example hydrochloric, hydrobromic,trifluoroacetic or maleic acid; or an alkali metal, for example sodium,alkaline earth metal or ammonium, for exampletetra(2-hydroxyethyl)ammonium, salt.

A suitable pharmaceutically-acceptable ester of a quinazoline of theinvention is, for example, an ester with an aliphatic alcohol of up to 6carbon atoms, for example a methyl, ethyl or tert-butyl ester.

It is to be understood that when R³ contains two carboxylic acidresidues, that is, when it is derived from, for example, aspartic orglutamic acid, a salt or ester may be a mono-acid-mono-salt or ester orit may be a di-salt or ester.

A preferred quinazoline of the invention has the formula stated abovewherein R¹ is methyl, ethyl, prop-2-enyl, prop-2-ynyl, methoxy,methylthio, phenyl, benzyl, fluoromethyl, difluoromethyl,trifluoromethyl, hydroxymethyl, aminomethyl, methoxymethyl,acetoxymethyl, methylthiomethyl, methylaminomethyl, dimethylaminomethylor acetamidomethyl; wherein R² is hydrogen, methyl, ethyl, propyl,prop-2-enyl, prop-2-ynyl, 2-hydroxyethyl, 2-methoxyethyl,2-mercaptoethyl, 2-methylthioethyl, 2-aminoethyl, 2-methylaminoethyl,2-dimethylaminoethyl, 2-bromoethyl or acetyl; wherein Ar is1,4-phenylene or thien-2,5-diyl which is unsubstituted or which bearsone substituent selected from chloro, methyl, methoxy or trifluoromethyland wherein R³ is such that R³ --NH₂ is L-alanine, L-glutamic acid orL-aspartic acid.

A further preferred quinazoline of the invention has the formula statedabove wherein R¹ is methyl, ethyl, isopropyl, cyclopropyl, cyclohexyl,methoxy, ethoxy, phenoxy, fluoro, chloro, hydroxy, mercapto,pyrimidin-2-ylthio, pyrimidin-2-ylthiomethyl, 2-hydroxyethoxy or2-methoxyethoxy; wherein R² is hydrogen, methyl, ethyl, prop-2-ynyl,3-hydroxypropyl, 3-methoxypropyl, 2-fluoroethyl, cyanomethyl, acetonyl,carboxymethyl or carbamoylmethyl; wherein Ar is 1,4-phenylene,thien-2,5-diyl, pyrid-2,5-diyl, pyrimidin-2,5-diyl, thiazol-2,5-diyl oroxazol-2,5-diyl which is unsubstituted or which bears one substituentselected from fluoro, chloro, cyano, nitro, hydroxy, amino or acetamidoand wherein R³ is such that R³ --NH₂ is L-glutamic acid, glycine,L-phenylalanine, L-serine, L-ornithine or L-aspartic acid.

An especially preferred quinazoline of the invention has the formulastated above wherein R¹ is methyl, ethyl, methoxy, fluoromethyl orhydroxymethyl; wherein R² is hydrogen, methyl, ethyl, propyl,prop-2-enyl, prop-2-ynyl or 2-hydroxyethyl; wherein Ar is 1,4-phenyleneor thien-2,5-diyl and wherein R³ is such that R³ --NH₂ is L-glutamicacid.

A further especially preferred quinazoline of the invention has theformula stated above wherein R¹ is methyl, methoxy, fluoromethyl orhydroxymethyl; wherein R² is hydrogen, methyl, ethyl, prop-2-ynyl,3-hydroxypropyl, 2-fluoroethyl or acetonyl; wherein Ar is 1,4-phenylene,thien-2,5-diyl, pyrid-2,5-diyl or 2-fluoro-1,4-phenylene and wherein R³is such that R³ --NH₂ is L-glutamic acid.

Specific particularly preferred quinazolines of the invention form thegroup of compounds:

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]benzoyl-L-glutamicacid,

N-p-[N-(2-ethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-o-fluorobenzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-fluorobenzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]-o-fluorobenzoyl-L-glutamicacid;

N-{5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl}-L-glutamicacid,

N-{5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-2-thenoyl}-L-glutamicacid,

N-{5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]picolinoyl}-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(2-fluoroethyl)amino]benzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-methoxy-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-hydroxymethyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid,

N-p-[N-(3,4-dihydro-2-hydroxymethyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]benzoyl-L-glutamicacid,

N-p-[N-(2-fluoromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid and

N-p-[N-(2-fluoromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]benzoyl-L-glutamicacid.

A quinazoline of the invention may be prepared by any process known tobe applicable to the preparation of chemically-related compounds.

A preferred process for the manufacture of a quinazoline of theinvention comprises the reaction of a compound of the formula: ##STR5##wherein R¹ has the meaning stated above, provided that when R¹ ishydroxyalkyl, aminoalkyl, alkylaminoalkyl or hydroxyalkoxy the hydroxyand amino groups are protected by conventional protecting group, R⁴ ishydrogen or a protecting group and Z is a displaceable group, with acompound of the formula:

    HNR.sup.2 --Ar--CONHR.sup.3

wherein R², Ar and R³ have the meanings stated above, provided that whenR² is hydroxyalkyl, mercaptoalkyl, aminoalkyl, alkylaminoalkyl orcarboxyalkyl, when there is an amino or hydroxy group in Ar or whenthere is an amino, hydroxy or carboxy group in R³, any mercapto, aminoand carboxy group is protected by a conventional protecting group andany hydroxy group may be protected by a conventional protecting group oralternatively any hydroxy group need not be protected; whereafter anyundesired protecting group in R¹, R², R³ and Ar is removed.

A suitable protecting group for a hydroxy group is, for example, anesterifying group, for example an acetyl or benzoyl group, which may beremoved by hydrolysis with a base, for example sodium hydroxide, orprovided that R¹ and R² do not contain an alkenyl or alkynyl group, theprotecting group may be, for example, an α-arylalkyl group, for examplea benzyl group, which may be removed by hydrogenation over a catalyst,for example palladium-on-charcoal.

A suitable protecting group for a mercapto group is, for example, anesterifying group, for example an acetyl group, which may be removed byhydrolysis with a base, for example sodium hydroxide.

A suitable protecting group for an amino group may be, for example, analkoxycarbonyl group, for example a tert-butyloxycarbonyl group whichmay be removed by treatment with an organic acid, for exampletrifluoroacetic acid; or it may be, for example, a benzyloxycarbonylgroup which may be removed by treatment with a Lewis acid, for exampleboron tris(trifluoroacetate).

A suitable alternative protecting group for a primary amino group is,for example, a phthaloyl group which may be removed by treatment with analkylamine, for example dimethylaminopropylamine or with hydrazine.

A suitable protecting group for a carboxy group may be an esterifyinggroup, for example a methyl or an ethyl group which may be removed byhydrolysis with a base, for example sodium hydroxide; or, for example atert-butyl group which may be removed by treatment with an organic acid,for example trifluoroacetic acid.

A suitable value for R⁴ when it is a protecting group is, for example, apivaloyloxymethyl group which may be removed by hydrolysis with a base,for example sodium hydroxide.

Z may be, for example, a halogeno or sulphonyloxy group, for example achloro, bromo, methanesulphonyloxy or toluene-p-sulphonyloxy group.

The protecting group for the various carboxy groups in R³ may beesterifying groups such as permit the product, after removal of theoptional protecting group R⁴ and of any undesired protecting groups inR¹, R², R³ or Ar, to fall within the definition of a quinazoline of theinvention. In such instance the carboxy protecting groups in R³ may beremoved or they may be retained. Alternatively a different protectinggroup may be used which will be removed.

A further preferred process for the manufacture of a quinazoline of theinvention comprises the reaction of an acid of the formula: ##STR6## ora reactive derivative thereof, with a compound of the formula R³ --NH₂,wherein R¹, R², R³, R⁴ and Ar have the meanings stated above and anymercapto, amino, alkylamino and carboxy group in R¹, R², R³ and Ar isprotected by a conventional protecting group, as stated above, and anyhydroxy group in R¹, R², R³ and Ar may be protected by a conventionalprotecting group, as stated above or alternatively any hydroxy groupneed not be protected; whereafter the protecting groups are removed byconventional means.

A suitable reactive derivative of an acid of the formula given above maybe, for example, an acyl halide, for example an acyl chloride formed bythe reaction of the acid and an inorganic acid chloride, for examplethionyl chloride; a mixed anhydride, for example an anhydride formed bythe reaction of the acid and a chloroformate such as isobutylchloroformate; an acyl azide, for example an azide formed by thereaction of the acid and an azide such as diphenylphosphoryl azide; orthe product of the reaction of the acid and a carbodiimide, for exampledicyclohexylcarbodiimide.

The carboxylic acid used as starting material may be obtained by thereaction of a compound of the formula ##STR7## wherein R¹, R⁴ and Z havethe meanings stated above, with a compound of the formula:

    HNR.sup.2 --Ar--CO.sub.2 R.sup.5

wherein R² and Ar have the meanings stated above and R⁵ is a protectinggroup which can be removed to provide a carboxylic acid.

R⁵ may be, for example, a methyl or an ethyl group which may be removedby hydrolysis with a base, for example sodium hydroxide or R⁵ may be,for example, a tert-butyl group which may be removed by cleavage with anorganic acid, for example trifluoroacetic acid.

The protecting group for the carboxy group in R⁵ may be, for example, anesterifying group which can be removed while the protecting group forany mercapto, amino, carboxy and hydroxy group in R¹, R² and Ar isretained.

A further preferred process for the manufacture of a quinazoline of theinvention, wherein R¹ is alkoxy, aryloxy or alkoxy of up to 3 carbonatoms which bears one or more substituents selected from hydroxy andalkoxy, comprises the reaction of a compound of the formula: ##STR8##wherein R¹ has the last-mentioned meaning stated above, provided thatwhen there is a hydroxy substituent in R¹ it is protected by aconventional protecting group, as stated above, and Z is a displaceablegroup, with a compound of the formula:

    HNR.sup.2 --Ar--CONHR.sup.3

wherein R², R³ and Ar have the meanings stated above, provided that whenR² is hydroxyalkyl, mercaptoalkyl, aminoalkyl, alkylaminoalkyl orcarboxyalkyl, when there is an amino or hydroxy group in Ar or whenthere is an amino, hydroxy or carboxy group in R³, any mercapto, aminoand carboxy group is protected by a conventional protecting group, asstated above, and any hydroxy group may be protected by a conventionalprotecting group, as stated above or alternatively any hydroxy groupneed not be protected; whereafter the protecting groups are removed byconventional means, as stated above and the R¹ group situated at the4-position of the quinazoline ring is removed by hydrolysis with a base,for example sodium hydroxide, to form a quinazoline of the invention.

A further preferred process for the manufacture of a quinazoline of theinvention, wherein R¹ is mercapto, alkylthio, pyridylthio orpyrimidinylthio, comprises the reaction of a quinazoline of the formula:##STR9## wherein R¹ is halogeno or halogenoalkyl and R², R³, R⁴ and Arhave the meanings stated above, provided that when R² is mercaptoalkyl,hydroxyalkyl, aminoalkyl, alkylaminoalkyl or carboxyalkyl, when there isan amino or hydroxy group in Ar or when there is an amino, hydroxy orcarboxy group in R³, any mercapto, amino, carboxy and hydroxy group maybe protected by a conventional protecting group, as stated above oralternatively any amino, carboxy and hydroxy group need not beprotected; with thiourea to provide a compound wherein R¹ is mercapto;or with an alkyl, pyridyl or pyrimidinyl thiol to provide a compoundwherein R¹ is alkylthio, pyridylthio, pyrimidinylthio, alkylthioalkyl,pyridylthioalkyl or pyrimidinylthioalkyl; whereafter the protectinggroups are removed by conventional means, as stated above.

A further preferred process for the manufacture of a quinazoline of theinvention, wherein R¹ is alkylthio, comprises the reaction of aquinazoline of the formula: ##STR10## wherein R¹ is mercapto and R², R³,R⁴ and Ar have the meanings stated above, provided that when R² ishydroxyalkyl, mercaptoalkyl, aminoalkyl, alkylaminoalkyl orcarboxyalkyl, when there is an amino or hydroxy group in Ar or whenthere is an amino, hydroxy or carboxy group in R³, any mercapto, amino,carboxy and hydroxy group may be protected by a conventional protectinggroup, as stated above or alternatively any amino, carboxyl and hydroxygroup need not be protected; with a base, for example ammonium hydroxideand the resultant thiolate salt is alkylated with an alkyl halide, forexample methyl iodide, to provide a compound wherein R¹ is alkylthio,for example methylthio; whereafter the protecting groups, if present,are removed by conventional means, as stated above.

As stated above a quinazoline of the invention possesses anti-tumouractivity and may itself be active thus or it may be a pro-drug which isconverted in vivo to an active compound. Preferred quinazolines of theinvention are 50 to 500 times more active than CB3717 in inhibiting thegrowth of the L1210 cell-line. L1210 is a mouse leukaemia cell linewhich can be grown in tissue culture (UK Patent Specification No.2065653B).

The quinazoline of the invention may be administered to a warm-bloodedanimal, including a human, in the form of a pharmaceutical compositionwhich comprises the quinazoline in association with apharmaceutically-acceptable diluent or carrier.

The composition may be in a form suitable for oral administration, as atablet or capsule, or, especially, for parenteral injection, as asterile solution, suspension or emulsion, or for topical administration,as an ointment or cream, or for rectal administration as a suppository.

The composition may contain, in addition to the quinazoline of theinvention, one or more other antitumour substances selected from, forexample, mitotic inhibitors, for example vinblastine; alkylating agents,for example cis-platin, carboplatin and cyclophosphamide; otherantimetabolites, for example 5-fluorouracil, cytosine arabinoside andhydroxyurea; intercalating antibiotics, for example adriamycin andbleomycin; enzymes, for example asparaginase; topoisomerase inhibitors,for example etoposide and biological response modifiers, for exampleinterferon.

The quinazoline will normally be administered to a warm-blooded animalat a dose within the range 50-5000 mg per square meter body area of theanimal.

The invention is illustrated but not limited by the following Examples:

The structures of all compounds of the invention were confirmed byproton magnetic resonance and mass spectroscopy and by elementalanalysis. Proton magnetic resonance spectra were determined using a JeolFX 90Q or a Bruker AM200 spectrometer operating at a field strength of200 MHz. Chemical shifts are reported in parts per million downfieldfrom tetramethylsilane as an internal standard (δ scale) and peakmultiplicities are shown thus: s, singlet; d, doublet; d of d's, doubletof doublets; t, triplet; m, multiplet. Fast-atom bombardment (FAB) massspectral data were obtained using a VG Analytical MS9 spectrometer andxenon gas and, where appropriate, either positive ion data or negativeion data were collected.

Column chromatography was performed using Merck Art 9385 silica gel.

EXAMPLE 1

A mixture of6-bromomethyl-3,4-dihydro-2-methyl-3-pivaloyloxymethylquinazolin-4-one(0.3 g), diethyl N-(p-prop-2-ynylaminobenzoyl)-L-glutamate (UK PatentSpecification No. 2065653B; 0.295 g), calcium carbonate (0.491 g) anddimethylformamide (10 ml) was stirred at 50° C. for 18 hours, cooled andfiltered through a filter-aid. The filtrate was evaporated to drynessand the residual oil was purified by chromatography on a silica gel(Merck 9385) column using a 3:1 v/v mixture of methylene chloride andethyl acetate as eluent.

A mixture of the product (0.306 g), ethanol (5 ml) and aqueous N-sodiumhydroxide solution (1.42 ml) was stirred at laboratory temperature for18 hours, acidified with acetic acid and aqueous 2N-hydrochloric acid(0.5 ml) was added. The mixture was centrifuged and the solid residuewas washed three times each with water and diethyl ether (10 ml eachtime) and dried. There was thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid (70 mg), m.p. (powder to glass) 165° C.

NMR Spectrum: (CD₃ SOCD₃) 2.0 (m, 2H, CH₂), 2.35 (broad t, 2H, CH₂ CO₂H), 2.35 (s, 3H, CH₃), 3.15 (t, 1H, C.tbd.CH, J=2 Hz), 4.3 (m, 3H, NHCHand CH₂ C.tbd.CH), 4.8 (s, 2H, CH₂ N), 6.83 (d, 2H, aromatic, J=9 Hz),7.52 (d, 1H, 8-H, J=9 Hz), 7.68 (d of d's, 1H, 7-H, J=2 and 9 Hz), 7.75(d, 2H, aromatic, J=9 Hz), 7.97 (d, 1H, 5-H, J=2 Hz), 8.18 (d, 1H, NH,J=8 Hz), 12.15 (broad s, 1H, NH).

Mass Spectrum: (positive ion FAB) m/e 477 (p+1).

Elemental Analysis: Found C, 58.9; H, 5.1; N, 10.9; C₂₅ H₂₄ N₄ O₆.2H₂ Orequires C, 58.6; H, 5.5; N, 10.9%.

The quinazolinone used as starting material was obtained as follows:

Sodium hydride (1.08 g) was added to a stirred suspension of3,4-dihydro-2,6-dimethylquinazolin-4-one (J. Indian Chem. Soc., 1962,39, 369; 3.0 g) in dimethylformamide (50 ml) and the mixture was stirredat laboratory temperature for 1 hour. A solution of chloromethylpivalate (3.36 g) in dimethylformamide (10 ml) was added and the mixturewas stirred at laboratory temperature for 18 hours and then poured intosaturated aqueous sodium chloride solution (200 ml). The mixture wasextracted four times with diethyl ether (50 ml each time) and thecombined extracts were dried and evaporated to dryness. The residue waspurified by chromatography on a silica gel column using a 9:1 v/vmixture of methylene chloride and ethyl acetate as eluent. The productwas crystallised from petroleum ether (b.p. 60°-80° C.) and there wasthus obtained3,4-dihydro-2,6-dimethyl-3-pivaloyloxymethyl-quinazolin-4-one (0.92 g),m.p. 98°-100° C.

A mixture of the above compound (0.92 g), N-bromosuccinimide (0.624 g),benzoyl peroxide (0.025 g) and carbon tetrachloride (50 ml) was heatedunder reflux for 2 hours, cooled and poured through a column of florisil(25 g). The column was eluted with carbon tetrachloride and the eluatewas evaporated to dryness. There was thus obtained as solid residue6-bromomethyl-3,4-dihydro-2-methyl-3-pivaloyloxymethyl-quinazolin-4-one(1.16 g), m.p. 144°-145° C.

EXAMPLE 2

The process described in Example 1 was repeated using diethylN-(p-ethylaminobenzoyl)-L-glutamate (British Journal of Cancer, 1979,40, 318) as starting material in place of the prop-2-ynylamino compound.There was thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]benzoyl-L-glutamicacid, m.p. 221°-225° C.

The process described in Example 1 was also repeated using6-bromomethyl-3,4-dihydro-3-pivaloyloxymethyl-2-trifluoromethylquinazolin-4-oneas starting material in place of the6-bromomethyl-2-methylquinazolin-4-one. There was thus obtainedN-p-[N-(3,4-dihydro-4-oxo-2-trifluoromethyl-quinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid, m.p. 110°-115° C.

3,4-Dihydro-6-methyl-2-trifluoromethylquinazolin-4-one used as startingmaterial was prepared by reacting trifluoroacetamide and2-amino-5-methylbenzoic acid using the method given in `The Chemistry ofHeterocyclic Compounds`, Volume 24, p 74.

EXAMPLE 3

A mixture of 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one (5.1 g),diethyl N-(p-methylaminobenzoyl)-L-glutamate (Journal of HeterocylicChemistry, 1975, 12, 1283; 6.7 g), 2,6-lutidine (7 ml) and drydimethylformamide (40 ml) was stirred at 80° C. under an atmosphere ofargon for 18 hours. The mixture was cooled, poured into water (300 ml)and extracted with ethyl acetate (4×150 ml). The combined extracts werewashed with water (2×200 ml), with a saturated aqueous sodium chloridesolution (2×100 ml), dried over magnesium sulphate, filtered andevaporated. The residue was purified by chromatography on a silica gelcolumn using ethyl acetate as eluent.

A mixture of the product (4.1 g), ethanol (25 ml) and aqueous N-sodiumhydroxide solution (24.3 ml) was stirred at laboratory temperature underan atmosphere of argon for 3 hours. The mixture was evaporated todryness, the residue was dissolved in de-ionised water and the solutionwas acidified to pH 2 by adding 2N-hydrochloric acid solution. Themixture was centrifuged and the solid residue was washed three timeswith water, diethyl ether and acetone (20 ml each time) and dried. Therewas thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]benzoyl-L-glutamicacid (containing 0.75 equivalents of water; 3 g), m.p. 254°-257° C.(decomposes).

NMR Spectrum: (CD₃ SOCD₃), 2.0 (m, 2H, CH₂), 2.35 (broad t, 2H, CH₂ CO₂H), 2.35 (s, 3H, CH₃), 3.12 (s, 3H, CH₃ N), 4.38 (m, 1H, NHCH, 4.78 (s,2H, CH₂ N), 6.77 (d, 2H, aromatic, J=9 Hz), 7.53 (d, 1H, 8-H, J=9 Hz),7.62 (d of d's, 1H, 7-H, J=2 and 9 Hz), 7.73 (d, 2H, aromatic, J=9 Hz),7.88 (d, 1H, 5-H, J=2 Hz), 8.15 (d, 1H, NH, J=8 Hz), 12.2 (s, 1H, NH);

Mass Spectrum: (positive ion FAB) m/e 453 (P+1).

Elemental Analysis: Found C, 59.1; H, 5.2; N, 11.9. C₂₃ H₂₄ N₄ O₆. 0.75H₂ O requires C, 59.3; H, 5.5; N, 12.0%.

The quinazolinone used as starting material was obtained as follows:

A mixture of 3,4-dihydro-2,6-dimethylquinazolin-4-one (20 g),N-bromosuccinimide (21.3 g), benzoyl peroxide (100 mg) and chloroform(600 ml) was heated to 50° C. for 6 hours during which time the mixturewas illuminated by the light from a 250 Watt light bulb. The mixture wascooled. The precipitated product was separated by filtration of themixture, washed with chloroform (2×50 ml) and dried. There was thusobtained 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one, m.p.>330°C.

EXAMPLE 4

The process described in Example 3 was repeated using the appropriate6-bromomethyl-3,4-dihydroquinazolin-4-one and the appropriate diethylp-aminobenzoyl-L-glutamate as starting materials. There were thusobtained the compounds described in the following table, the structuresof which were confirmed by proton magnetic resonance and massspectroscopy and by elemental analysis.

                                      TABLE I                                     __________________________________________________________________________     ##STR11##                                                                    EXAMPLE 4                                                                     Compound No                                                                           R.sup.1  (Note)                                                                            R.sup.2   m.p.   x                                       __________________________________________________________________________     1      methyl   (1) H         197-201° C.                                                                   1                                        2      methyl   (1) prop-2-enyl                                                                             188° C. (dec.)                                                                1.5                                      3      methyl   (1) 3-hydroxypropyl                                                                        >300° C. (dec.)                                                                1.2                                      4      methyl   (1) 2-fluoroethyl                                                                           207-210° C.                                                                   1.2                                      5      methyl   (2) 2-hydroxyethyl                                                                         >300° C. (dec.)                                                                1.5                                      6      methyl   (2) 2-methoxyethyl                                                                          248° C. (dec.)                                                                1.0                                      7      methyl   (2) 3-methoxypropyl                                                                         260° C. (dec.)                                                                1.0                                      8      methyl   (2) acetonyl  155-157° C.                                                                   1.0                                      9      ethyl    (3) prop-2-ynyl                                                                             150-157° C.                                                                   0.5                                     10      ethyl    (3) H         156-166° C.                                                                   2                                       11      isopropyl                                                                              (3) prop-2-ynyl                                                                             148-150° C.                                                                   2                                       12      phenyl   (4) prop-2-ynyl                                                                             170° C.                                                                       0.5                                     13      difluoromethyl                                                                         (4) prop-2-ynyl                                                                             135-140° C.                                                                   1                                       14      hydroxymethyl                                                                          (5) prop-2-ynyl                                                                             137-143° C.                                                                   2                                       15      hydroxymethyl                                                                          (5) prop-2-enyl                                                                             150-160° C.                                                                   1                                       16      hydroxymethyl                                                                          (5) ethyl     140-150° C.                                                                   1                                       17      hydroxymethyl                                                                          (5) methyl    194-197° C.                                                                   1                                       18      hydroxymethyl                                                                          (5) 2-hydroxyethyl                                                                          150-155° C.                                                                   1                                       19      hydroxymethyl                                                                          (5) 2-fluoroethyl                                                                           215-222° C.                                                                   0.5                                     20      acetamidomethyl                                                                        (6) prop-2-ynyl                                                                             229-240° C.                                                                   1.5                                     21      chloro   (7) prop-2-ynyl                                                                             156-160° C.                                                                   3                                       __________________________________________________________________________    Note (1): The appropriate diethyl  .sub.--p-aminobenzoyl-L-glutamate          was obtained as described in the literature (Journal of                       Medicinal Chemistry, 1985, 28, 1468 or the European Journal of                Cancer, 1981, 17, 11).                                                        Note (2): The required diethyl glutamate was prepared by                      reaction of diethyl  .sub.--p-aminobenzoyl-L-glutamate with the               alkylating agents 2-acetoxyethyl bromide,                                     2-methoxyethyl bromide, 3-methoxypropyl bromide and 1-bromoacetone            in an analogous process to that described in the literature                   (Journal of Medicinal Chemistry 1985, 28, 1468).                              Note (3): The required quinazolinones were obtained using the                 method described in `The Chemistry of Heterocyclic Compounds`                 Volume 24, page 74 with propionamide and isobutyramide                        respectively instead of acetamide as the starting material.                   Note (4): The required quinazolinones were prepared by the                    method described in the literature (J. Amer. Chem. Soc. 1946, 68,             1299 and UK Patent Specification No. 1410178).                                Note (5): 2-Acetoxymethyl-3,4-dihydro-6-methylquinazolin-4-one                (Dissertationes Pharmaceuticae et Pharmacolgicae 1968, 20, 29)                and the appropriate diethyl  .sub.--p-aminobenzoyl-L-glutamate were           taken through the process described in Example 3. Basic                       hydrolysis cleaved the glutamate esters and the acetoxy group.                Note (6): 2-Chloromethyl-3,4-dihydro-6-methylquinazolin-4-one                 (Dissertationes Pharmaceuticae et Pharmacologicae 1968, 20, 29)               was treated with a saturated aqueous solution of amonia at                    laboratory temperature for 20 hours. The solvent was removed                  and the product was acetylated to give 2-acetamidomethyl-3,4-                 dihydro-6-methylquinazolin-4-one which was used as the starting               material in the sequence described in Example 3.                              Note (7): 2-Chloro-3,4-dihydro-6-methylquinazolin-4-one was                   obtained as described in U.S. Pat. No. 4,085,213.                         

EXAMPLE 5

A mixture of 6-bromomethyl-2-fluoromethyl-3,4-dihydroquinazolin-4-one(0.62 g), di-tert-butyl N-(p-prop-2-ynylaminobenzoyl)-L-glutamate (1.2g, prepared by reaction of di-tert-butyl N-p-aminobenzoyl-L-glutamate,known from the Journal of Medicinal Chemistry, 1985, 28, 1468, withprop-2-ynyl bromide using the method described in the European Journalof Cancer 1981, 17, 11), 2,6-lutidine (1.5 g) and dry dimethylformamide(20 ml) was stirred at 60° C. for 18 hours under an atmosphere of argon.The mixture was cooled, the solvent was evaporated and the residual oilwas purified by chromatography on a florisil column using a 2:1 v/vmixture of methylene chloride and ethyl acetate as eluent.

A mixture of the product (0.6 g), trifluoroacetic acid (2 ml) andchloroform (6 ml) was stirred at laboratory temperature for 4 hours. Themixture was poured into diethyl ether (40 ml) and stirred for 10minutes. The precipitated solid was separated by filtration of themixture and the solid was washed with ether (3×10 ml) and dried. Therewas thus obtainedN-p-[N-(2-fluoromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid as a dihydrate, trifluoroacetic acid salt, (0.3 g), m.p. 126°-131°C.

NMR Spectrum: (CD₃ SOCD₃) 2.0 (m, 2H, CH₂), 2.3 (t, 2H, CH₂ CO₂ H, J=6.5Hz), 3.18 (t, 1H, C.tbd.CH, J=2 Hz), 4.15 (m, 3H, NHCH and CH₂C.tbd.CH), 4.8 (s, 2H, CH₂ N), 5.27 (d, 2H, FCH₂, J=47 Hz), 6.84 (d, 2H,aromatic, J=9 Hz), 7.66 (d, 1H, 8-H, J=9 Hz), 7.75 (m, 3H, aromatic and7-H), 8.04 (d, 1H, 5-H, J=2 Hz), 8.21 (d, 1H, NH, J=8 Hz).

Mass Spectrum: (negative ion FAB) m/e 493 (P-1).

Elemental Analysis: Found C, 50.5; H, 4.1; N, 9.2. C₂₅ H₂₃ FN₄ O₆.CF₃CO₂ H.2H₂ O requires C, 50.3; H, 4.3; N, 8.7%.

The quinazolinone used as starting material was obtained as follows:

A mixture of 2-amino-5-methylbenzoic acid (20 g) and fluoroacetamide (40g) was heated to 120° C. for 1 hour, to 140° C. for 90 minutes and to180° C. for 90 minutes. The mixture was cooled to room temperature andthe residue was purified by chromatography on a silica gel column usinga 1:1 v/v mixture of methylene chloride and ethyl acetate as eluent.

A mixture of the 2-fluoromethyl-3,4-dihydro-6-methylquinazolin-4-one (2g) so obtained, N-bromosuccinimide (1.8 g), benzoyl peroxide (10 mg) andchloroform (50 ml) was heated to reflux for 4 hours, cooled andevaporated. The product,6-bromomethyl-2-fluoromethyl-3,4-dihydroquinazolin-4-one was usedwithout further purification.

The process described above was repeated using di-tert-butylN-(p-ethylaminobenzoyl)-L-glutamate with ethyl iodide using the methoddescribed above in the first paragraph of this Example) in place of theprop-2-ynylamino compound. There was thus obtainedN-p-[N-(2-fluoromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]benzoyl-L-glutamicacid as a hemi-trifluoroacetic acid salt, m.p. 162°-167° C.

EXAMPLE 6

A mixture of 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one(prepared as described in Example 3 above, 0.38 g), diethylN-(2-fluoro-4-methylaminobenzoyl)-L-glutamate (prepared by the reactionof diethyl N-(4-amino-2-fluorobenzoyl)-L-glutamate, known from UK PatentSpecification No. 2175903, with methyl iodide using the method describedin the European Journal of Cancer, 1981, 17, 11; 0.7 g), powderedcalcium carbonate (0.3 g) and dry dimethylformamide (2.7 ml) was stirredat 100° C. for 7 hours. The mixture was evaporated and the residue waspurified by chromatography on a silica gel column using a 20:1 v/vmixture of methylene chloride and ethanol as eluent.

A mixture of the product (0.54 g), ethanol (10 ml), water (10 ml) andaqueous N-sodium hydroxide solution (6.2 ml) was stirred at laboratorytemperature for 7 hours. The mixture was concentrated to a volume ofapproximately 5 ml, filtered and acidified to pH 3 by adding2N-hydrochloric acid solution. The precipitated solid was isolated bycentrifugation, washed with water (4×30 ml) and dried. There was thusobtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-o-fluorobenzoyl-L-glutamicacid monohydrate (0.41 g), m.p. 224°-226° C.

EXAMPLE 7

The process described in Example 6 was repeated using the appropriatediethyl L-glutamate as starting material. There were thus obtained thecompounds described in the following table, the structures of which wereconfirmed by proton magnetic resonance and mass spectroscopy and byelemental analysis.

                                      TABLE II                                    __________________________________________________________________________     ##STR12##                                                                    EXAMPLE 7                                                                     Compound No.                                                                          R.sup.2 (Note)                                                                             Ar      x   m.p.                                         __________________________________________________________________________    1       H       (1)                                                                                 ##STR13##                                                                            1   190-194° C.                           2       ethyl   (2)                                                                                 ##STR14##                                                                            0.75                                                                              214-217° C.                           3       prop-2-ynyl                                                                           (2)                                                                                 ##STR15##                                                                            1   228-230° C.                           4       HOCH.sub.2 CH.sub.2                                                                   (2)                                                                                 ##STR16##                                                                            0   190-196° C.                           5       FCH.sub.2 CH.sub.2                                                                    (2)                                                                                 ##STR17##                                                                            0.7 220-225° C.                           6       H.sub.2 NCOCH.sub.2                                                                   (3)                                                                                 ##STR18##                                                                            1   170-185° C.                           7       prop-2-ynyl                                                                           (4)  thien-2,5-diyl                                                                        0.5 215-225° C.                           8       methyl  (5)  thien-2,5-diyl                                                                        1   180-184° C.                           9       ethyl   (5)  thien-2,5-diyl                                                                        0.75                                                                              162-167° C.                           10      n-propyl                                                                              (5)  thien-2,5-diyl                                                                        2   184-185° C.                           11      methyl  (6)                                                                                 ##STR19##                                                                            0.75                                                                              203-205° C.                           12      prop-2-ynyl                                                                           (7)                                                                                 ##STR20##                                                                            0.5 240-248° C.                           13      methyl  (8)                                                                                 ##STR21##                                                                            2.5 200-204° C.                           __________________________________________________________________________    Note (1): The preparation of diethyl  --N-(4-amino-2-                         fluorobenzoyl)-L-glutamate is described in UK Patent                          Specification No. 2175903.                                                    Note (2): The appropriate diethyl glutamate was prepared by the               reaction of diethyl  --N-(4-amino-2-fluorobenzoyl)-L-glutamate (UK            Patent Specification No. 2175903) with ethyl iodide, propargyl                bromide, 2-acetoxyethyl bromide or 2-fluoroethyl bromide                      respectively, using the method described in the Journal of                    Medicinal Chemistry, 1985, 28, 1468.                                          Note (3): Diethyl  --N- .sub. --p-( --N-cyanomethylamino)- .sub.--o-fluoro    benzoyl-L-                                                                    glutamate was prepared by the method given in Note (2) above                  using chloroacetonitrile as the alkylating agent. During the                  final step of the process described in Example 6 the  --N-                    cyanomethylamino group was hydrolysed to an  --N-                             carbamoylmethylamino group.                                                   Note (4): 2,6-Lutidine was used in place of calcium carbonate.                Diethyl  --N-[5-(prop-2-ynylamino)-2-thenoyl]-L-glutamate used as             starting material was obtained as follows:-                                   Pyridine (1.5 g) was added to a mixture of 5-nitro-2-                         thenoyl chloride (3 g) and diethyl L-glutamate hydrochloride                  (4.2 g) in toluene (50 ml). The mixture was stirred at                        laboratory temperature for 72 hours, poured into water (200 ml)               and extracted with ethyl acetate (3 × 100 ml). The combined             extracts were washed with water (2 × 500 ml) and with a                 saturated aqueous sodium chloride solution (1 × 50 ml), dried           over magnesium sulphate, filtered and evaporated to dryness.                  The residue was purified by chromatography on a silica gel                    column using a 9:1 v/v mixture of methylene chloride and ethyl                acetate as eluent.                                                            A mixture of the product (3.4 g), ethanol (15 ml) and                         an aqueous solution of sodium dithionite (6.2 g of the dihydrate              salt in 30 ml of water) was stirred at 50° C. for 1 hour, poured       into water (100 ml) and extracted with ethyl acetate (3 ×               50 ml). The combined extracts were washed with a saturated                    aqueous sodium chloride solution (1 × 50 ml), dried over                magnesium sulphate, filtered and evaporated to dryness. The                   residue was purified by chromatography on a silica gel column                 using a 7:3 v/v mixture of methylene chloride and ethyl acetate               as eluent. There was thus obtained diethyl 5-amino-2-thenoyl-L-               glutamate (1 g).                                                              A mixture of the product (1 g), propargyl bromide                             (0.66 g of an 80% solution in toluene), 2,6-lutidine (0.5 g)                  and dimethylformamide (25 ml) was stirred at 50° C. for 24 hours,      cooled, poured into water (25 ml) and extracted with ethyl                    acetate (3 × 50 ml). The combined extracts were washed with             water (2 × 25 ml) and with a saturated aqueous sodium chloride          solution (25 ml), dried over magnesium sulphate, filtered and                 evapoarted to dryness. The residue was purified by                            chromatography on a silica gel column using a 7:3 v/v mixture of              methylene chloride and ethyl acetate as eluent. There was thus                obtained diethyl  --N-[5-(prop-2-ynylamino)-2-thenoyl]-L-glutamate            (0.6 g).                                                                      Note (5): 2,6-Lutidine was used in place of calcium carbonate.                The appropriate diethyl L-glutamate was obtained using diethyl                5-amino-2-thenoyl-L-glutamate as starting material in the                     process described in the last paragraph of note (4) above except              that methyl iodide, ethyl iodide and n-propyl iodide                          respectively were used in place of propargyl bromide.                         Note (6): Diethyl  --N-[5-(methylamino)picolinoyl]-L-glutamate                used as starting material was prepared as follows:- A mixture                 of methyl 5-( --N-tert-butoxycarbonyl- --N-methylamino)picolinate             (prepared using the method described in the Journal of Medicinal              Chemistry, 1980, 23, 1405 except that methyl iodide was used in               place of 3-trifluoromethylbenzyl chloride; 1.13 g), aqueous N-                sodium hydroxide solution (8.5 ml), water (21 ml) and ethanol                 (15 ml) was stirred at laboratory temperature for 16 hours. The               mixture was concentrated to a volume of 5 ml, acidified to pH 4               with 2N-hydrochloric acid solution and extracted with ethyl                   acetate (2 × 40 ml). The combined extracts were dried over              magnesium sulphate, filtered and evaporated to give 5-( --N-tert-             butoxycarbonyl-  --N-methylamino)picolinic acid (0.8 g).                      A mixture of the acid (0.74 g), oxalyl chloride                               (0.38 ml), methylene chloride (8 ml) and dimethylformamide was                stirred at laboratory temperature for 45 minutes and evaporated               to dryness. A solution of the residue in methylene chloride                   (10 ml) was added to a mixture of diethyl L-glutamate                         hydrochloride (0.77 g), 2.6-lutidine (0.65 ml) and methylene                  chloride (10 ml). The mixture was stirred at laboratory                       temperature for 16 hours, washed with N-hydrochloric acid                     solution, a saturated aqueous sodium bicarbonate solution and a               saturated aqueous sodium chloride solution, dried over magnesium              sulphate, filtered and evaporated to dryness. The residue was                 purified by chromatography on a silica gel column using a 10:1                v/v mixture of methylene chloride and ethyl acetate as eluent.                There was thus obtained diethyl  --N-[5-( --N-tert-butoxycarbonyl- --N-       methylamino)picolinyl]-L-glutamate (0.48 g).                                  A mixture of this ester and trifluoroacetic acid                              (10 ml) was stirred at 0° C. for 1 hour and evaporated to              dryness.                                                                      The residue was washed with diethyl ether and dried. There was                thus obtained as a gum diethyl  --N-[5-(methylamino)picolinyl]-L-             glutamate (0.4 g) which was used without further purification.                Note (7): The process described in Note (6) was repeated                      except that methyl 5-[ --N-tert-butoxycarbonyl- --N-(prop-2-                  ynyl)amino]picolinate (prepared using the method described in                 the Journal of Medicinal Chemistry 1980, 23, 1405, except that                propargyl bromide was used in place of 3-trifluoeomethylbenzyl                chloride) was used in place of the corresponding                              methylaminopicolinate. There was thus obtained as a gum diethyl                --N-{5-[ --N-(prop-2-ynyl)amino]picolinyl}-L-glutamate.                      Note (8): The mono-sodium salt of the glutamic acid was                       obtained. Diethyl  --N-[6-(methylamino)nicotinoyl]-L-glutamate                used as starting material was prepared as follows:-                           A mixture of 6-chloronicotinic acid (8.27 g) oxalyl                           chloride (5.63 ml), methylene chloride (70 ml) and                            dimethylformamide (0.25 ml) was stirred at laboratory                         temperature for 16 hours and evaporated to dryness. A solution                of the residue in methylene chloride (100 ml) was added to a                  mixture of diethyl L-glutamate hydrochloride (13.6 g),                        triethylamine (22 ml) and methylene chloride (100 ml). The                    mixture was stirred at laboratory temperature for 16 hours,                   washed with water (100 ml), dried over sodium sulphate, filtered              and evaporated. The residue was purified by chromatography on a               silica gel column using a 4:1 v/v mixture of methylene chloride               and ethyl acetate as eluent. There was thus obtained diethyl  --N-            (6-chloronicotinoyl)-L-glutamate (17 g), as an oil.                           A mixture of this product (0.53 g),  --N-benzyl- --N-                         methylamine (0.49 ml) and  --N-methylpyrrolidin-2-one (2 ml) was              stirred and heated to 100° C. for 16 hours under an atmosphere of      argon. The mixture was evaporated and the residue was                         partitioned between ethyl acetate and a saturated aqueous sodium              bicarbonate solution. The organic layer was dried over sodium                 sulphate, filtered and evaporated. The residue was purified by                chromatography on a silica gel column using a 4:1 v/v mixture of              methylene chloride and ethyl acetate as eluent. There was thus                obtained diethyl  --N-[6-( --N-benzyl- --N-methylamino)nicotinoyl]-L-         glutamate (0.57 g) as an oil.                                                 A mixture of this product (0.3 g), trifluoroacetic                            acid (1 ml), palladium-on-charcoal catalyst (10%, 0.05 g) and                 ethanol (2 ml) was stirred at 60° C. for 2 hours. The mixture was      filtered and the filtrate was evaporated. The residue was                     partitioned between ethyl acetate and a saturated aqueous sodium              bicarbonate solution. The organic layer was dried over sodium                 sulphate, filtered and evaporated. There was thus obtained                    diethyl  --N-[6-(methylamino)nicotinoyl]-L-glutamate (0.22 g) which           was used without further purification.                                    

EXAMPLE 8

The process described in the first paragraph of Example 3 was repeatedexcept that 6-bromomethyl-2,4-dimethoxyquinazoline was used in place of6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one.

A mixture of the product (1.8 g), ethanol (20 ml), aqueous N-sodiumhydroxide solution (26.9 ml) and water (20 ml) was stirred at 60° C. for16 hours. The mixture was evaporated on a rotary evaporator to a volumeof approximately 10 ml, filtered and acidified to pH 3 by adding2N-hydrochloric acid solution. The mixture was centrifuged and the solidresidue was washed with water (4×30 ml) and dried. There was thusobtainedN-p-[N-(3,4-dihydro-2-methoxy-4-oxoquinazolin-6-ylmethyl)-N-methylamino]benzoyl-L-glutamicacid (containing 2.5 equivalents of water; 0.55 g), m.p. 240°-245° C.

NMR Spectrum: (CD₃ SOCD₃) 2.0 (m, 2H, CH₂), 2.32 (t, 2H, CH₂ CO₂ H),3.09 (s, 3H, CH₃ N), 3.93 (s, 3H, CH₃ O), 4.45 (m, 1H, NHCH), 4.75 (s,2H, CH₂ N), 6.77 (d, 2H, aromatic, J=9 Hz), 7.44 (d, 1H, 8-H, J=9 Hz),7.55 (d of d's, 1H, 7-H, J=2 and 9 Hz), 7.74 (d, 2H, aromatic, J=9 Hz),7.84 (d, 1H, 5-H, J=2 Hz), 8.17 (d, 1H, NH, J=8 Hz).

Mass Spectrum: (negative ion FAB) m/e 467 (P-1).

Elemental Analysis: Found C, 53.8; H, 4.8; N, 10.7. C₂₃ H₂₄ N₄ O₇.2.5 H₂O requires C, 53.8; H, 5.65; N, 10.9%.

The bromomethylquinazoline used as starting material was obtained asfollows:

A mixture of 2,4-dimethoxy-6-methylquinazoline (8.2 g),N-bromosuccinimide (7.9 g), benzoyl peroxide (0.19 g) and carbontetrachloride (200 ml) was heated to reflux for 2 hours. The warmsolution was filtered and the filtrate was evaporated to give6-bromomethyl-2,4-dimethoxyquinazoline (11.7 g), m.p. 138°-143° C.

EXAMPLE 9

The process described in Example 8 was repeated except that theappropriate diethyl or di-tert-butyl L-glutamate was used in place ofdiethyl N-(p-methylaminobenzoyl)-L-glutamate. There were thus obtainedthe compounds described in the following table, the structures of whichwere confirmed by proton magnetic resonance and mass spectroscopy and byelemental analysis.

                                      TABLE III                                   __________________________________________________________________________     ##STR22##                                                                    EXAMPLE 9                                                                     Compound No                                                                           R.sup.2  (Note)                                                                            Ar      x   m.p.                                         __________________________________________________________________________    1       H            1,4-phenylene                                                                         1   150-160° C.                           2       ethyl        1,4-phenylene                                                                         1   140-146° C.                           3       prop-2-ynyl  1,4-phenylene                                                                         1   155-165° C.                           4       prop-2-enyl  1,4-phenylene                                                                         1   130-134° C.                           5       2-hydroxyethyl                                                                             1,4-phenylene                                                                         1.25                                                                              150-175° C.                           6       3-hydroxypropyl                                                                            1,4-phenylene                                                                         1.5 145-155° C.                           7       2-fluoroethyl                                                                              1,4-phenylene                                                                         1.25                                                                              141-145° C.                           8       carboxymethyl                                                                          (1) 1,4-phenylene                                                                         2   165-185° C.                           9       2-aminoethyl                                                                           (2) 1,4-phenylene                                                                         0.75                                                                              217-220° C.                           10      ethyl                                                                                       ##STR23##                                                                            0.5 140-145° C.                           11      ethyl        thien-2,5-diyl                                                                        1.25                                                                              132-135°  C.                          __________________________________________________________________________    Note (1): Diethyl  --p-(carboethoxymethylamino)benzoyl-L-glutamate            used in the preparation of this compound has been described                   (Journal of Medicinal Chemistry, 1985, 28, 1468).                             Note (2): Di-tert-butyl  --N-[ .sub.--p-(2-phthalimidoethyl)aminobenzoyl]-    L-glutamate was reacted with 6-bromomethyl-2,4-                               dimethoxyquinazoline using the process described in the first                 paragraph of Example 3. A mixture of the product (2.1 g), 3-                  dimethylaminopropylamine (2.12 ml), di-isopropylethylamine                    (0.98 ml) and methanol (18 ml) was heated to reflux for 11 hours              and evaporated to dryness. The residue was purified by                        chromatography on a silica gel column using a 20:1 v/v mixture                of methylene chloride and methanol as eluent. There was thus                  obtained di-tert-butyl  --N- .sub.--p-[ --N-(2-aminoethyl)- --N-(2,4-         dimethoxyquinazolin-6-ylmethyl)amino]benzoyl-L-glutamate                      (0.7 g). This di-ester was hydrolysed using the conditions                    described in the second paragraph of Example 8 to provide                     --N- .sub.--p-                                                                [ --N-(2-aminoethyl)- --N-(3,4-dihydro-2-methoxy-4-oxoquinazolin-6-           ylmethyl)amino]benzoyl-L-glutamic acid.                                       The di-tert-butyl ester used as starting material was                         obtained as follows:-                                                         A mixture of di-tert-butyl  -- N-( .sub.--p-aminobenzoyl-L-glutamate          (Journal of Medicinal Chemistry 1985, 28, 1468; 5.1 g),  --N-(2-              bromoethyl)phthalimide (20.4 g), 2,6-lutidine (9.4 ml) and  --N, --N-         dimethylacetamide (20 ml) was heated to 100° C. for 18 hours           under                                                                         an atmosphere of argon. The mixture was cooled, poured into                   aqueous N-sulphuric acid solution (110 ml) and extracted with                 ethyl acetate (3 × 70 ml). The combined extracts were washed            with a saturated aqueous sodium chloride solution (3 × 50 ml),          dried over magnesium sulphate, filtered and evaporated. The                   residue was purified by chromatography on a silica gel column                 using a 10:1 v/v mixture of methylene chloride and ethyl acetate              as eluent. There was thus obtained di-tert-butyl  --N-[ .sub.--p-(2-          phthalimidoethyl)aminobenzoyl]-L-glutamate (5.3 g),                           m.p. 157-158° C.                                                   

EXAMPLE 10

The process described in Example 8 was repeated using diethylN-[p-(prop-2-ynyl)aminobenzoyl)-L-glutamate and the appropriate2,4-dialkoxy- or 2,4-diaryloxy-6-bromomethyl-quinazoline. There was thusobtained the compounds described in the following table, the structuresof which were confirmed by proton magnetic resonance and massspectroscopy and by elemental analysis.

                                      TABLE IV                                    __________________________________________________________________________     ##STR24##                                                                    EXAMPLE 10                                                                    Compound No                                                                              R.sup.1    (Note)                                                                            x     m.p.                                          __________________________________________________________________________    1          ethoxy     (1) 2     134-136° C.                            2          2-methoxyethoxy                                                                          (2) 1     134-140° C.                            3          2-hydroxyethoxy                                                                          (3) 2     145-149° C.                            4          phenoxy    (4) 3     159-164° C.                            __________________________________________________________________________    Note (1): The bromomethyl compound used as starting                           material was obtained as follows:-                                            A mixture of 2,4-dichloro-6-methylquinazoline (3 g)                           and a solution of sodium ethoxide [made by adding sodium metal                (1.07 g) to ethanol (100 ml)] was heated to reflux for 4 hours,               cooled, poured into a saturated aqueous sodium chloride solution              (100 ml) and extracted with ethyl acetate (3 × 90 ml). The              combined extracts were washed with water, dried over magnesium                sulphate, filtered and evaporated to dryness. The residue was                 purified by chromatography on a silica gel column using a 10:1                v/v mixture of methylene chloride and ethyl acetate as eluent.                There was thus obtained 2,4-diethoxy-6-methylquinazoline                      (1.9 g), m.p. 60-62° C., which was converted to the 6-bromomethyl      derivative using the process described in the last paragraph of               Example 8.                                                                    Note (2): The bromomethyl compound used as starting material                  was obtained using the process described above, starting from                 2,4-dichloro-6-methylquinazoline but using the sodium salt of                 2-methoxyethanol instead of sodium ethoxide.                                  Note (3): The bromomethyl compound used as starting material                  was obtained as follows:-                                                     Ethylene glycol (30 ml) was added to sodium hydride                           (2.4 g of a 50% dispersion in oil which was washed with hexane                under an atmosphere of argon). A solution of 2,4-dichloro-6-                  methylquinazoline (2.06 g) in dimethylformamide (5 ml) was added              and the mixture was stirred at 100° C. for 16 hours, cooled,           poured onto water (100 ml) and extracted with ethyl acetate (3 ×        200 ml). The combined extracts were washed with a saturated                   aqeous sodium chloride solution, dried over magnesium sulphate,               filtered and evaporated. A mixture of the product (2 g),                      anhydrous pyridine (20 ml) and benzoyl chloride (1.9 ml) was                  stirred at laboratory temperature for 16 hours, poured into                   water (100 ml) and extracted with methylene chloride (3 ×               70 ml). The combined extracts were washed with a saturated                    aqueous sodium bicarbonate solution (100 ml) and with water                   (50 ml), dried over magnesium sulphate, filtered and evaporated.              The residue was purified by chromatography on a silica gel                    column using a 20:1 v/v mixture of methylene chloride and ethyl               acetate as eluent. There was thus obtained 2,4-di-(2-                         benzoyloxyethoxy)-6-methylquinazoline (1.3 g) which was                       converted to the 6-bromomethyl derivative using the process                   described in the last paragraph of Example 8.                                 The process described in Example 8 was then repeated                          and the final aqueous basic hydrolysis was used to removed                    the benzoyl protecting groups.                                                Note (4): The bromomethyl compound used as starting material                  was obtained as follows:-                                                     A mixture of a solution of sodium phenoxide in molten                         phenol (obtained on the addition of sodium metal (0.8 g)                      to molten phenol (30 g) at 80° C.) and 2,4-dichloro-6-                 methylquinazoline (3.2 g) was heated to 180° C. for 1 hour. The        warm mixture was poured into water (200 ml), an aqueous                       10N-sodium hydroxide solution (5 ml) was added followed by                    glacial acetic acid to bring the acidity of the mixture to pH 6.              The solid was filtered off, washed with water, dissolved in                   methylene chloride and dried over magnesium sulphate. The                     solution was passed through a silica gel column using more                    methylene chloride as eluent. There was thus obtained 2,4-                    diphenoxy-6-methylquinazoline (4.6 g), m.p. 184-185° C., which         was                                                                           converted to the 6-bromomethyl derivative using the process                   described in the last paragraph of Example 8.                             

EXAMPLE 11

Diphenylphosphoryl azide (0.44 g) and triethylamine (0.67 ml) were addedsuccessively to a mixture ofp-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoicacid (as its trifluoroacetic acid salt; 0.5 g), L-alanine ethyl ester(as its hydrochloride salt; 0.27 g) and dimethylformamide (20 ml) whichwas cooled in an ice-bath to 0° C. The mixture was stirred at 0° C. for5 hours and at laboratory temperature for 48 hours, poured into amixture of ice and water (100 ml) and centrifuged. The solid residue waswashed with water (3×10 ml) and dried. The residue was purified bychromatography on a silica column using a 24:1 v/v mixture of methylenechloride and ethanol as eluent.

A mixture of the product (0.11 g), ethanol (4 ml), water (4 ml) andaqueous N-sodium hydroxide solution (0.64 ml) was stirred at laboratorytemperature for 2 hours, acidified to pH 3 with aqueous0.2N-hydrochloric acid solution and centrifuged. The solid residue waswashed with water (5×10 ml) and dried. There was thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-alanine(0.08 g), as a monohydrate, m.p. 165°-170° C.

NMR Spectrum: (CD₃ SOCD₃) 1.32 (d, 3H, CH₃ ; J=7 Hz), 2.31 (s, 3H, CH₃),3.18 (t, 1H, C.tbd.CH, J=2 Hz), 4.3 (m, 3H, NHCH and CH₂ C.tbd.CH), 4.78(s, 2H, CH₂ N), 6.83 (d, 2H, aromatic, J=9 Hz), 7.52 (d, 1H, 8-H, J=8.5Hz), 7.68 (d of d's, 1H, 7-H, J=2 and 8.5 Hz), 7.72 (d, 2H, aromatic,J=9 Hz), 7.96 (d, 1H, 5-H, J=2 Hz), 8.21 (d, 1H, NH, J=6.5 Hz), 12.13(s, 1H, NH);

Mass Spectrum: (negative ion FAB) m/e 418 (P-1).

Elemental Analysis: Found C, 63.0; H, 5.3; N, 12.3. C₂₃ H₂₂ N₄ O₄.H₂ Orequires C, 63.3; H, 5.5; N, 12.8%.

The process described in Example 11 was repeated using L-phenylalanineethyl ester, L-serine methyl ester and L-aspartic acid dimethyl esterrespectively in place of alanine ethyl ester. There were thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-phenylalanineas a monohydrate, m.p. 152°-155° C., the corresponding benzoyl-L-serine,as a hemi-hydrate m.p. 200°-204° C., and the correspondingbenzoyl-L-aspartic acid (containing 1.25 equivalents of water), m.p.180°-190° C. (decomposes).

The process described in the first paragraph of Example 11 was alsorepeated using N⁵ -benzyloxycarbonyl-L-ornithine tert-butyl ester inplace of alanine ethyl ester.

Boron tris(trifluoroacetate) (1 ml of a 1 molar solution intrifluoroacetic acid) was added to a solution of the product (0.1 g) intrifluoroacetic acid (1 ml) which had been cooled to -10° C. The mixturewas stirred at 5° C. for 3 hours, methanol (2 ml) was added and themixture was evaporated. The residue was purified by chromatography on apreparative thin-layer chromatography plate using a 4:1 v/v mixture ofethanol and an aqueous ammonia solution (concentrated) as solvent. Therewas thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-ornithine,as a monohydrate (15 mg), m.p. 210°-215° C. (decomposes).

N-p-[N-(3,4-Dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoicacid used as starting material was obtained as follows:

A mixture of tert-butyl p-aminobenzoate (Synth. Commun., 1984, 14, 921;10.5 g), propargyl bromide (7.3 ml of an 80% solution in toluene),potassium carbonate (7.5 g) and N,N-dimethylacetamide (85 ml) was heatedto 50° C. for 24 hours, cooled, filtered and evaporated. The residue waspurified by chromatography on a silica gel column using a 6:1 v/vmixture of hexane and ethyl acetate as eluent.

A mixture of the product (7.3 g);6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one (prepared asdescribed in Example 3 above; 8 g), calcium carbonate (3.2 g) anddimethylformamide (100 ml) was stirred at laboratory temperature for 65hours, filtered and evaporated. The residue was purified bychromatography on a silica gel column using ethyl acetate as eluent.

The mixture of the product (2.5 g) and trifluoroacetic acid (25 ml) wasstirred at laboratory temperature for 10 minutes and evaporated to givethe p-aminobenzoic acid as its trifluoroacetic acid salt (2.5 g).

EXAMPLE 12

A mixture of 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one (1.24g), methyl N-[p-(prop-2-ynyl)aminobenzoyl]glycine (prepared as describedin the Journal of Medicinal Chemistry, 1986, 29, 1117; 1.2 g), calciumcarbonate (0.5 g) and dimethylformamide (12 ml) was stirred atlaboratory temperature for 72 hours, filtered and evaporated. Theresidue was purified by chromatography on a column of silica gel using a9:1 v/v mixture of ethyl acetate and methanol as eluent.

A portion of the product (0.17 g) was hydrolysed under basic conditionsusing the process described in the second paragraph of Example 11. Therewas thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoylglycine(0.09 g; containing 1.5 equivalents of water) m.p. 240°-250° C.(decomposes).

EXAMPLE 13

The process described in the second paragraph of Example 3 was repeatedexcept that diethylN-p-[N-(3,4-dihydro-2-methylthio-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamatewas used as starting material. There was thus obtainedN-p-[N-(3,4-dihydro-2-methylthio-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid (containing 0.75 equivalents of water), m.p. 157°-163° C.

The starting material was obtained as follows:

A mixture of diethylN-p-[N-(2-chloro-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate(obtained using the process described in Example 4; 0.75 g), thiourea(0.125 g), formic acid (0.05 ml) and ethanol (20 ml) was heated toreflux for 15 minutes, cooled and evaporated to dryness. The residue waspurified by chromatography on a silica gel column using a 10:3 v/vmixture of methylene chloride and ethyl acetate as eluent. There wasthus obtained diethylN-p-[N-(4-oxo-1,2,3,4-tetrahydro-2-thioxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate,m.p. 92°-94° C.

A mixture of this product (0.19 g), water (12.8 ml), ethanol (9.5 ml)and an aqueous ammonia solution (3.2 ml of a solution of specificgravity of 0.88 g ml⁻¹) was stirred at laboratory temperature for 10minutes. Methyl iodide (0.13 ml) was added and the mixture was stirredfor 1 hour. The precipitated solid was filtered off, washed with a 1:1v/v mixture of water and ethanol and dried. There was thus obtaineddiethylN-p-[N-(3,4-dihydro-2-methylthio-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate(0.16 g; containing 0.75 equivalents of water), m.p. 230°-233° C.

Alternatively the product described in the first paragraph aboveconcerned with the production of starting materials may be hydrolysedwith base using the process described in the second paragraph of Example3. There was thus obtainedN-p-[N-(4-oxo-1,2,3,4-tetrahydro-2-thioxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamicacid as a monohydrate, m.p. 161°-166° C.

EXAMPLE 14

The process described in the second paragraph of Example 3 was repeatedexcept that diethylN-p-{N-[3,4-dihydro-4-oxo-2-(pyrimidin-2-ylthio)quinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino}benzoyl-L-glutamatewas used as starting material. There was thus obtainedN-p-{N-[3,4-dihydro-4-oxo-2-(pyrimidin-2-ylthio)quinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino}benzoyl-L-glutamicacid (containing 0.5 equivalents of water), m.p. 143°-147° C.

The diethyl p-aminobenzoyl-L-glutamate used as starting material wasobtained as follows:

A mixture of diethylN-p-[N-(2-chloro-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate(obtained using the process described in Example 4; 0.35 g),2-mercaptopyrimidine (0.21 g) and N-methylpyrrolid-2-one (5 ml) wasstirred at laboratory temperature for 16 hours, poured into water (20ml) and extracted with ethyl acetate (3×20 ml). The combined extractswere washed with water, dried over magnesium sulphate, filtered andevaporated. The residue was purified by chromatography on a silica gelcolumn using a 3:2 v/v mixture of methylene chloride and ethyl acetateas eluent. There was thus obtained diethylN-p-{N-[3,4-dihydro-4-oxo-2-(pyrimidin-2-ylthio)quinazolin-6-ylmethyl]-N-(prop-2-ynyl)-amino}benzoyl-L-glutamate(0.19 g), as a monohydrate, m.p. 163°-165° C.

EXAMPLE 15

A mixture of diethylN-p-[N-2-chloromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate(0.56 g), 2-mercaptopyrimidine (0.11 g), sodium hydride (0.047 g of a50% dispersion in oil which was washed with hexane) anddimethylformamide (10 ml) was stirred at laboratory temperature for 16hours, poured into water (50 ml) and extracted with ethyl acetate (4×25ml). The combined extracts were washed with water (2×25 ml), dried overmagnesium sulphte, filtered and evaporated. The residue was purified bychromatography on a column of silica gel using ethyl acetate as eluent.

The product was hydrolysed with base using the process described in thesecond paragraph of Example 3. There was thus obtainedN-p-{N-[3,4-dihydro-4-oxo-2-(pyrimidin-2-ylthiomethyl)quinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino}benzoyl-L-glutamicacid (0.32 g; containing 1.5 equivalents of water), m.p. 151°-153° C.

The diethyl L-glutamate used as starting material was obtained asfollows:

The process described in the paragraph in Example 3 which is concernedwith the preparation of starting materials was repeated except that2-chloromethyl-3,4-dihydro-6-methylquinazolin-4-one (DissertationesPharmaceuticae et Pharmacologicae 1968, 20, 29) was used in place of3,4-dihydro-2,6-dimethylquinazolin-4-one. There was thus obtained6-bromomethyl-2-chloromethyl-3,4-dihydroquinazolin-4-one.

The process described in the first paragraph of Example 3 was repeatedexcept that the 6-bromomethyl-2-chloromethyl-3,4-dihydroquinazolin-4-oneand diethyl N-p-(prop-2-ynyl)aminobenzoyl-L-glutamate where used asstarting materials. There was thus obtained diethylN-p-[N-(2-chloromethyl-3,4-dihydro-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]benzoyl-L-glutamate.

EXAMPLE 16

A mixture of 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-One (5.1 g),diethyl N-p-(prop-2-ynyl)amino-o-trifluoromethylbenzoyl-L-glutamate (1.1g), magnesium oxide (0.12 g) and N,N-dimethylacetamide (30 ml) wasstirred and heated to 80° C. for 19 hours. The mixture was cooled,poured onto ice (100 ml) and extracted with ethyl acetate (3×200 ml).The combined extracts were washed with water (2×100 ml), dried oversodium sulphate, filtered and evaporated. The residue was purified bychromatography on a silica gel column using a 50:1 v/v mixture ofmethylene chloride and methanol as eluent. There was thus obtaineddiethylN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]-o-trifluoromethylbenzoyl-L-glutamate(0.96 g), m.p. 191° C.

A mixture of a portion of this product (0.49 g), ethanol (15 ml), water(15 ml) and aqueous N-sodium hydroxide solution (2.5 ml) was stirred atlaboratory temperature for 17 hours. The mixture was filtered and thefiltrate was acidified to pH 4 by adding N-hydrochloric acid solution.The mixture was centrifuged and the solid residue was washed three timeswith water and dried. There was thus obtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-(prop-2-ynyl)amino]-o-trifluoromethylbenzoyl-L-glutamicacid (0.38 g, as hemihydrate), m.p. 197° C.

The diethyl glutamate used as starting material was obtained as follows:

A mixture of 4-nitro-2-trifluoromethylbenzonitrile (J. Amer. Chem. Soc.1954, 76, 1051; 5.6 g), glacial acetic acid (20 ml) and sulphuric acid(concentrated, 30 ml) was stirred and heated to 130° C. for 45 minutes.The mixture was cooled, poured once ice (100 ml) and extracted withethyl acetate (3×150 ml). The combined extracts were washed with aqueous0.05N-hydrochloric acid solution, dried over sodium sulphate, filteredand evaporated. The residue was purified by chromatography on a silicagel column using ethyl acetate as eluent. There was thus obtained4-nitro-2-trifluoromethylbenzamide (5.16 g), m.p. 192° C.

A mixture of this product (1.82 g), water (50 ml), sodium hydroxide (2g) and hydrogen peroxide (30%, 10 ml) was stirred and heated to 70° C.for 4 hours during which time one further portion of sodium hydroxide (2g) and two further portions of hydrogen peroxide (30%, 10 ml each time)were added. The mixture was heated to 70° C. for 3 days, cooled,acidified with aqueous N-hydrochloric acid solution and extracted withethyl acetate (3×100 ml). The combined extracts were washed with aqueous0.05N-hydrochloric acid acid solution, dried over sodium sulphate,filtered and evaporated to leave, as a light brown solid,4-nitro-2-trifluoromethylbenzoic acid (1.76 g), m.p. 128°-129° C. (J.Amer. Chem. Soc., 1954, 76, 1051; m.p. 137°-140° C.).

A mixture of this product (0.79 g), toluene (50 ml) and thionyl chloride(2 ml) was heated to reflux for 5 hours, cooled and evaporated. Asolution of the residue in methylene chloride (50 ml) was added to astirred mixture of diethyl L-glutamate hydrochloride (0.68 g),triethylamine (0.75 g) and methylene chloride (100 ml) which was cooledto 4° C. The mixture was stirred at laboratory temperature for 2 hours,washed with water (4×100 ml), dried over sodium sulphate, filtered andevaporated. The residue was purified by chromatography on a silica gelcolumn using a 9:1 v/v mixture of methylene chloride and ethyl acetateas eluent. There was thus obtained ethylN-p-nitro-o-trifluoromethylbenzoyl-L-glutamate (1.23 g) m.p. 105° C.(recrystallised from ethanol solution).

After repetition of the above reactions on a larger scale a mixture ofthis product (12.5 g), ethanol (1 litre) and palladium-on-charcoalcatalyst (10%, 1 g) was stirred under an atmosphere of hydrogen untilthe calculated volume of hydrogen had been consumed. The mixture wasfiltered and evaporated to leave an oil which crystallised on standing.There was thus obtained diethylN-p-amino-o-trifluoromethylbenzoyl-L-glutamate (11.6 g), m.p. 95° C.

A mixture of this product (10.2 g), propargyl bromide (as an 80%solution in toluene, 8.5 g), potassium carbonate (7.2 g) and drydimethylformamide (150 ml) was stirred and heated to 100° C. for 100minutes. The mixture was cooled, poured onto ice (100 ml) and extractedwith ethyl acetate (3×200 ml). The combined extracts were washed withwater (2×200 ml), dried over sodium sulphate, filtered and evaporated.The residue was purified by chromatography on a silica gel column usinga 2:1 v/v mixture of petroleum ether (b.p. 60°-80° C.) and ethyl acetateas eluent. There was thus obtained diethylN-p-(prop-2-ynyl)amino-o-trifluoromethylbenzoyl-L-glutamate (7.3 g),m.p. 91° C.

EXAMPLE 17

The process described in Example 3 was repeated using6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one (prepared asdescribed in Example 3) as the appropriate quinazoline; the appropriatediethyl L-glutamate and the appropriate organic or inorganic base in thefirst step. There were thus obtained the compounds described in thefollowing table, the structures of which were confirmed by protonmagnetic resonance and mass spectroscopy and by elemental analysis.

                                      TABLE V                                     __________________________________________________________________________     ##STR25##                                                                    Example 17                                                                    Compound No.                                                                          R.sup.2                                                                              Note                                                                              Ar         x  m.p.                                         __________________________________________________________________________    1       ethyl  (1)                                                                                ##STR26## 1  152-157° C.                           2       prop-2-ynyl                                                                          (2)                                                                                ##STR27## 2.5                                                                              175-180° C.                           3       ethyl  (3)                                                                                ##STR28## 1  211° C. (dec)                         4       prop-2-ynyl                                                                          (4)                                                                                ##STR29## 1  156-160° C.                           5       prop-2-ynyl                                                                          (5)                                                                                ##STR30## 1  210° C. (dec)                         6       ethyl  (6)                                                                                ##STR31## 1  201-207° C.                           7       prop-2-ynyl                                                                          (6)                                                                                ##STR32## 0.5                                                                              162-164° C.                           __________________________________________________________________________    Note (1): Diethyl  --N(4-ethylamino-2-methoxybenzoyl)-L-glutamate             was obtained using the process described in the last three                    paragraphs of the portion of Example 16 which is concerned with               the preparation of starting materials except that 2-methoxy-4-                nitrobenzoic acid (Journal of the Chemical Society, 1917, 111,                232) was used in place of 4-nitro-2-trifluoromethylbenzoic acid               and ethyl iodide was used in place of propargyl bromide.                      Note (2): Diethyl  --N[2-methoxy-4-(prop-2-ynyl)aminobenzoyl]-L-              glutamate was obtained using the process described in the last                three paragraphs of the portion of Example 16 which is concerned              with the preparation of starting materials except that 2-methoxy-             4-nitrobenzoic acid (Journal of the Chemical Society, 1917, 111,              232) was used in place of 4-nitro-2-trifluoromethylbenzoic acid.              Note (3): Diethyl  --N-(2-acetamido-4-ethylaminobenzoyl)-L-glutamate          was obtained using the process described in the last three                    paragraphs of the portion of Example 16 which is concerned with               the preparation of starting materials except that 2-acetamido-4-              nitrobenzoic acid (Journal of the Chemical Society, 1925, 127,                1795, was used in place of 4-nitro-2-trifluoromethylbenzoic acid              and ethyl iodide was used in place of propargyl bromide.                      Note (4) Naphthalene-1,8-diamine was used as the base in place                of 2,6-lutidine. Diethyl  --N-[3-fluoro-4-(prop-2-ynyl)amino-                 benzoyl]-L-glutamate was obtained using the process described in              the last three paragraphs of the portion of Example 16 which is               concerned with the preparation of starting materials except that              3-fluoro-4-nitrobenzoic acid (Journal of the American Chemical                Society, 1944, 66, 1631) was used in place of 4-nitro-3-                      trifluoromethylbenzoic acid.                                                  Note (5): Diethyl  --N-[2-acetoxy-4-(prop-2-ynyl)aminobenzoyl] L-             glutamate was obtained using the process described in the last                three paragraphs of the portion of Example 16 which is concerned              with the preparation of starting materials except that 2-acetoxy-             4-nitrobenzoic acid [obtained by the reaction of 2-hydroxy-4-                 nitrobenzoic acid (The Dictionary of Organic Compounds, Volume                3, page 3169; Chapman and Hall, 1982) and acetic anhydride at                 laboratory temperature] was used in place of 4-nitro-3-                       trifluorobenzoic acid.                                                        The conditions of the last step of the process                                described in Example 3, that is, the hydrolysis under basic                   conditions of the appropriate diethyl glutamate, resulted in the              hydrolysis of the 2-acetoxy group. There was thus obtained                    --N- .sub.--p-                                                                [ --N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)- --N-(prop-2-         ynyl)amino]- .sub.--o-hydroxybenzoyl-L-glutamic acid.                         Note (6): Diethyl  --N-[2-chloro-4-(prop-2-ynyl)aminobenzoyl]-L-              glutamate was prepared from diethyl  --N-(4-amino-2-chlorobenzoyl)-           L-glutamate (Journal of Medicinal Chemistry, 1986, 29, 468). The              corresponding 4-ethylaminobenzoyl-L-glutamate was prepared from               the 4-amino derivative using the method described in the European             Journal of Cancer, 1981, 17, 11.                                              __________________________________________________________________________

EXAMPLE 18

A mixture ofp-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-nitrobenzoicacid (2.5 g), oxalyl chloride (0.93 g), tetrahydrofuran (200 ml) anddimethylformamide (1 drop) was stirred at laboratory temperature for 18hours and evaporated. A solution of the residue in tetrahydrofuran (200ml) was added to a stirred mixture of diethyl L-glutamate hydrochloride(1.77 g), triethylamine (10 ml) and tetrahydrofuran (25 ml). The mixturewas stirred at laboratory temperature for 2 hours, washed with water(2×50 ml), with a saturated aqueous sodium chloride solution (50 ml),dried over magnesium sulphate, filtered and evaporated. The residue waspurified by chromatography on a silica gel column using a 10:1 v/vmixture of ethyl acetate and methanol as eluent. There was thus obtaineddiethylN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-nitrobenzoyl-L-glutamate(0.64 g).

A mixture of this product (0.64 g) and an aqueous N-sodium hydroxidesolution (10 ml) was stirred at laboratory temperature for 2 hours thenacidified to pH 4 by the addition of an aqueous 2N-hydrochloric acidsolution. The mixture was centrifuged and the solid residue was washedwith water (4×10 ml) and acetone (10 ml) and dried. There was thusobtainedN-p-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-nitrobenzoyl-L-glutamicacid as a monohydrate (0.13 g), m.p. 192°-200° C. (decomposes)

The benzoic acid used as starting material was obtained as follows:

A mixture of methyl-p-amino-o-nitrobenzoate (The Dictionary of OrganicCompounds, Volume 1, page 285; Chapman and Hall, 1982; 1 g), ethyliodide (0.8 g), 2,6-lutidine (2.7 g) and dimethylformamide (5 ml) wasstirred and heated to 80° C. for 18 hours, cooled and evaporated. Amixture of the residue, 2,6-lutidine (2.7 g),6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one (1.3 g) anddimethylformamide (10 ml) was stirred at 85° C. for 5 hours, cooled,poured into water (100 ml) and extracted with ethyl acetate (3×70 ml).The combined extracts were washed with a saturated aqueous sodiumchloride solution (70 ml), dried over magnesium sulphate, filtered andevaporated. The residue was purified by chromatography on a silica gelcolumn using a 50:1 v/v mixture of ethyl acetate and methanol as eluent.There was thus obtained methylp-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-nitrobenzoate(0.35 g).

A mixture of this product (0.35 g) and aqueous N-sodium hydroxidesolution (10 ml) was stirred at laboratory temperature for 4 hours. Themixture was acidified to pH 4 by the addition of aqueous 2N-hydrochloricacid solution. The precipitated solid was separated by filtration anddried. There was thus obtainedp-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]-o-nitrobenzoicacid (0.3 g).

EXAMPLE 19

The process described in Example 3 was repeated using2-acetoxymethyl-6-bromomethyl-3,4-dihydroquinazolin-4-one (prepared fromthe 6-methyl compound, which is described in note (5) of Example 4,using the process described in the portion of Example 3 concerned withthe preparation of starting materials) in place of6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one and, in turn, diethylN-(5-methylamino-2-theonyl)-L-glutamate and diethylN-(5-ethylamino-2-thenoyl)-L-glutamate (both of which were prepared asdescribed in note (5) of Example 7) in place of diethylN-(p-methylaminobenzoyl)-L-glutamate. There were thus obtainedN-5-[N-(3,4-dihydro-2-hydroxymethyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino-2-thenoyl-L-glutamicacid, as its monohydrate, m.p. 180°-190° C. andN-{5-[N-(3,4-dihydro-2-hydroxymethyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino-2-thenoyl}-L-glutamicacid, as its monohydrate, m.p. 148°-153° C.

EXAMPLE 20

The process described in Example 11 was repeated except that2-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]thiazole-5-carboxylicacid was used in place of the benzoic acid and diethyl L-glutamate, asits hydrochloride salt, was used in place of L-alanine ethyl ester.There was thus obtainedN-2-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]thiazole-5-carbonyl-L-glutamicacid as its hemi-hydrate, m.p. 160°-170° C.

The thiazole-5-carboxylic acid used as starting material was obtained asfollows:

A mixture of 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one(prepared as described in Example 3; 10 g), anhydrous ethylamine (7.9ml) and acetonitrile (250 ml) was stirred rapidly at laboratorytemperature for 4 hours. The mixture was evaporated to dryness, theresidue was dissolved in water, filtered and the filtrate wasevaporated. The residue was triturated in acetone to giveN-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamine, as itshydrobromide salt (8.5 g), m.p. >260° C.

A mixture of this product (6.1 g), benzoyl isothiocyanate (2.75 ml) andacetone (25 ml) was stirred and heated to 50° C. for 2 hours. Themixture was poured into water (250 ml) and the product was filtered offand dried. A mixture of this solid, aqueous hydrochloric acid(concentrated, 80 ml) and isopropanol (48 ml) was stirred and heated to100° C. for 30 minutes. The mixture was evaporated and the residue wastriturated in ethyl acetate to giveN-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylthiourea (5.3g), m.p. 186°-187° C.

A mixture of the thiourea (4.67 g), ethyl formylchloroacetate (Archivder Pharmazie, 1953, 286, 494; 2.55 g) and dimethylformamide (25 ml) wasstirred and heated to 100° C. for 1 hour. The mixture was cooled,filtered and the filtrate was evaporated. The residue was partitionedbetween methylene chloride and a saturated aqueous sodium bicarbonatesolution. The organic solution was dried over sodium sulphate, filteredand evaporated. The residue was purified by trituration in ethyl acetateto give ethyl2-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-ethylamino]thiazole-5-carboxylate(1.37 g), m.p. 188°-192° C.

A mixture of this ester (1.3 g) and an aqueous N-sodium hydroxidesolution (10.5 ml) was heated to 48° C. for 1 hour. The mixture wascooled and acidified to pH 4 by the addition of an aqueous2N-hydrochloric acid solution. The gummy precipitate was isolated bycentrifugation and triturated in water to give the thiazole-5-carboxylicacid (1.05 g) used as starting material above.

EXAMPLE 21

The process described in Example 3 was repeated except that6-bromomethyl-1,2,3,4-tetrahydroquinazolin-2,4-dione was used in placeof 6-bromomethyl-3,4-dihydro-2-methylquinazolin-4-one and diethylN-(p-ethylaminobenzoyl)-L-glutamate was used in place of thep-methylaminobenzoyl derivative. There was thus obtainedN-p-[N-ethyl-N-(1,2,3,4-tetrahydro-2,4-dioxoquinazolin-6-ylmethyl)amino]benzoyl-L-glutamicacid as a hemi-hydrate, m.p. 205°-211° C.

The 6-bromomethyl-1,2,3,4-tetrahydroquinazolin-2,4-dione used asstarting material was obtained from1,2,3,4-tetrahydro-6-methylquinazolin-2,4-dione (Journal of HeterocyclicChemistry, 1984, 21, 5) using the method described in the secondparagraph of the portion of Example 5 which is concerned with thepreparation of starting materials.

What we claim is:
 1. A method for producing an antitumor effect in awarm-blooded animal in need of such treatment, which comprisesadministering to said animal an effective amount of at least onequinazoline of the formula ##STR33## wherein R¹ is alkyl, cycloalkyl,alkenyl, alkynyl, alkoxy or alkylthio each of up to 6 carbon atoms;aryl, aryloxy or arylalkyl each of up to 10 carbon atoms; halogeno,hydroxy, mercapto, pyridylthio or pyrimidinylthio; alkyl of up to 3carbon atoms which bears one, two or three halogeno sustituents or whichbears one or two substituents selected from hydroxy, amino, pyridylthio,pyrimidinylthio, alkoxy, alkanoyloxy, alkylthio, alkylamino,dialkylamino and alkanoylamino each of up to 6 carbon atoms and aroyloxyand aroylamino each of up to 10 carbon atoms; or alkoxy of up to 3carbon atoms which bears one or two substituents selected from hydroxyand alkoxy of up to 6 carbon atoms;wherein R² is hydrogen, alkyl,alkenyl, alkynyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl,alkylthioalkyl, halogenoalkyl, cyanoalkyl, aminoalkyl, alkylaminoalkyl,dialkylaminoalkyl, alkanoylalkyl, carboxyalkyl, carbamoylalkyl oralkanoyl each of up to 6 carbon atoms or aroylalkyl of up to 10 carbonatoms; wherein Ar is phenylene, napthylene or heterocyclene which isunsubstituted or which bears one or two substituents selected fromhalogeno, phenyl, cyano, nitro, hydroxy, amino and carbamoyl and alkyl,alkoxy, halogenoalkyl, alkanoylamino, alkylthio and alkoxycarbonyl eachof up to 6 carbon atoms; and wherein R³ is such that R³ --NH₂ isL-aspartic acid, L-glutamic acid, L-alanine, L-phenylalanine, L-serine,glycine, L-ornithine, L-2-aminobutyric acid or a poly-L-glutamic acid ofthe formula ##STR34## wherein m is an integer from 1 to 10; or apharmaceutically acceptable salt or ester thereof.